let's briefly talk about how to use the step one plus to do qpcr so we hit the screen the power button is this blue button in the bottom left that will take a second to get fired up this is very similar to the Quant Studio systems it does have an extra die and um if you check it's currently not posted right now but if you check this cork board that's right here by the time you see this there will be a die guide that says which dies are calibrated in each machine and which ones are compatible I'm here um you can make shortcuts of protocols so for example this is already set up to do a standard and a fast attack man protocol make sure you pick the correct one we can open one of these you could modify if you click each stage you can then either add a stage or you can go to options and there's a ton of options in terms of the rate at which you're ramping or the amount of time you spend in each cycle when you're collecting each data point and then if you were going to add a melt curve at a certain stage which it looks like there may not be one configured in this protocol currently you could add a stage and then add another curve um although actually that little c in there I bet that's the Melt curve right there we hit save when we're done editing our protocol and we're just going to overwrite the existing experiment and this is the protocol that we set from our shortcuts if you picked a different shortcut that wasn't shown here once you save it it will appear here and go into this is our experiment that we're about to run I'm gonna open this this is a lid so you can put this aside it's just a blank plate make sure that the plate you're using is similar and fits apply bio systems inside of this machine put this aside and you can place your assay plate into the wells if you have a deeper well plate we have this insert which adds a little bit of extra height that allows a deeper well plate to be put in so that you don't basically you have more space for your plate to rest we close that in and then we would hit start run to begin so very simple I mean when you're done it will you'll have the option to put in a USB key to export your your experimental results we go to the main menu um and then there's this button that says collect results uh you need to insert a USB key for that work but then that will guide you through the process of putting your data onto again when you're done take out your plate put the lid back in and then turn the power off on the machine that's about it let us know if you have any questions